CN115286530B - Chiral hydrogel material functionalized by halogenation effect, preparation method and application thereof - Google Patents

Chiral hydrogel material functionalized by halogenation effect, preparation method and application thereof Download PDF

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CN115286530B
CN115286530B CN202211054919.XA CN202211054919A CN115286530B CN 115286530 B CN115286530 B CN 115286530B CN 202211054919 A CN202211054919 A CN 202211054919A CN 115286530 B CN115286530 B CN 115286530B
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刘进营
赵伟利
冯传良
卢会杰
张健
张言言
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Abstract

The invention belongs to the field of supermolecular chemistry, and relates to a chiral hydrogel material with a halogenation effect functionalization, a preparation method and application thereof. By introducing different halogen atoms and changing substitution positions or side chain-hydrophilic hydrophobicity of the halogen atoms, the space conformation of the gel factor is changed, and the chiral hydrogel material with different spiral chirality and halogenation effect functionalization is constructed by single chiral molecules. The gelator self-assembles through physical interactions of hydrogen bonding, aromatic stacking, fluorine hydrogen bonding, and halogen bonding. The invention obviously improves the defects of poor biocompatibility, unstable structure and difficult operation of the existing system, increases the physicochemical property of halogen atoms, and realizes the chiral uniformity and biological function diversity of the supermolecular hydrogel.

Description

Chiral hydrogel material functionalized by halogenation effect, preparation method and application thereof
Technical Field
The invention belongs to the field of supermolecular chemistry, and relates to a chiral hydrogel material with a halogenation effect functionalization, a preparation method and application thereof.
Background
Chirality is one of the most important chemical signals in nature, and has important effects on the replication of genetic information, transcription, formation of higher structures of proteins, and protein functionalization. Plays a decisive role in maintaining the normal functions of human tissue cells. Intermolecular forces such as pi-pi stacking, hydrogen bonding, electrostatic interactions, and van der Waals forces are key determinants of natural systems for the transfer and amplification of molecular chirality of the basic unit of an organism into supramolecular chiral structures. The construction of the chiral micro-nano structure and the exploration of the intrinsic mechanism of the chiral influence cell behavior have application value in the field of biological medicine and have great significance in discussing the chiral origin of natural life. In recent years, scholars at home and abroad research on construction of molecular chiral structures and supermolecular chiral assembly strategies, and develop a plurality of chiral assemblies with good biocompatibility and diversified functions. Nevertheless, there are a number of critical scientific issues that need to be addressed with regard to the construction of supramolecular chiral assemblies still in the early exploration phase. Among them, the existing chiral assemblies are mostly formed by molecular modification, the change of assembly environment (pH, solvent and temperature) and co-assembly to mediate non-covalent interaction to realize the amplification of molecular chiral signals and control supermolecular chiral inversion, so that the existing chiral structure is far from the original chiral structure in complexity and functionality, and chiral assemblies with uniform chirality are difficult to obtain. Thus, new forces and assembly strategies are needed to drive more complex supramolecular materials.
Halogenation effect refers to the introduction of a halogen atom (F, cl, br or I) into an organic compound, which changes a certain property of the compound or material due to its unique chemical nature. Wherein the covalent halogen atoms in the organic halide may act as non-covalent interactions between the lewis acid and neutral or negatively charged lewis base, an intermolecular weak interaction resembling hydrogen bonds. The fluorine atom has unique intrinsic properties such as strong electronegativity and small atomic radius, so that the fluorine atom can participate in not only the formation of halogen bonds, but also the formation of various covalent interactions such as the formation of hydrogen bonds as a receptor of the hydrogen bonds. The non-covalent interaction of halogen atoms has wide application in many fields such as crystal engineering, functional material design, drug design, etc., but the halogenation effect has few reports on controlling the influence of supermolecule chiral structure.
Disclosure of Invention
Aiming at the technical problems, the invention provides a chiral hydrogel material with a functional halogenation effect, and a preparation method and application thereof. The invention changes the space conformation of chemical molecules by utilizing the halogenation effect of substitution of different halogen atoms or the halogenation effect of different substitution positions of the same halogen atom, so that the chirality of the chiral hydrogel material with the halogenation effect functionalized is correspondingly changed, and the spiral fiber structure with different chiralities constructed by single enantiomer is obtained. When used for cell adhesion proliferation, the composition has outstanding effect. The preparation method provided by the invention does not need complex reaction, is easy for large-scale production, and the purity of the obtained product is high; the adhesion behavior of the chiral nanofiber hydrogel regulatory cells is carried out in a physiological environment, so that the chiral nanofiber hydrogel regulatory cell is convenient to operate and has practical application value.
In order to achieve the above purpose, the technical scheme of the invention is realized as follows:
the chiral hydrogel material with the functional halogenation effect is formed by constructing the chiral hydrogel material with different spiral chirality and the functional halogenation effect by a single chiral molecule through introducing different halogen atoms and changing the substitution positions of the halogen atoms or changing the side chain-hydrophilic hydrophobicity, and meanwhile, the gel factors are self-assembled through the physical interactions of hydrogen bonds, aromatic stacking effects, fluorine hydrogen bonds and halogen bonds.
Further, the gel factor is an L-phenylalanine derivative, and the structural formula of the chiral hydrogel material with the functional halogenation effect is shown as I, II or III:
Figure GDA0004212129440000021
Figure GDA0004212129440000022
Figure GDA0004212129440000031
wherein, -F represents any one of ortho-position, meta-position and para-position of F substitution; r1 is->
Figure GDA0004212129440000032
R is
Figure GDA0004212129440000033
X is any one of fluorine, chlorine, bromine or iodine.
Further, the preparation method of the chiral hydrogel material functionalized by the halogenation effect comprises the following steps:
(1) When the chiral hydrogel material with the functional halogenation effect is of a structural formula shown as I, the preparation steps are as follows:
a1, dissolving fluorine-substituted Boc-L-phenylalanine and 1-Hydroxybenzotriazole (HOBT) with dichloromethane, cooling in an ice bath, adding diglycolamine and N, N-Diisopropylethylamine (DIPEA), and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, and continuing the room temperature reaction for 12 hours; sequentially washing with saturated citric acid solution, saturated sodium bicarbonate solution, saturated sodium chloride solution, spin drying, and recrystallizing with ethyl acetate and petroleum ether to obtain Boc-L-NF-phe with structural formula of
Figure GDA0004212129440000034
Represents that the F substitution position is any one of ortho position, meta position or para position;
a2, adding the Boc-L-NF-phe obtained in the step a1 into a mixed solution of dichloromethane and trifluoroacetic acid, reacting for 3 hours at room temperature, removing Boc-protecting groups, and completely removing the solution and the residual trifluoroacetic acid of the L-NF-phe by rotary evaporation of the solution to obtain the L-NF-phe trifluoroacetate, wherein the structural formula is as follows
Figure GDA0004212129440000035
Represents that the F substitution position is any one of ortho position, meta position or para position;
a3, dissolving the L-NF-phe trifluoroacetate obtained in the step a2 in dichloromethane, cooling in an ice bath, and sequentially addingAdding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 24h, filtering the colloid after the reaction, washing dichloromethane, ethanol and deionized water in sequence, and drying the sample to obtain a white solid target product I, wherein the structural formula is as follows
Figure GDA0004212129440000041
The F substitution position is any one of ortho-position, meta-position or para-position.
Further, when the chiral hydrogel material functionalized by the halogenation effect has a structural formula shown as I, the fluorine atom substituted Boc-L-phenylalanine and diglycolamine in the step a1 undergo an amide condensation reaction in the presence of N, N-Diisopropylethylamine (DIPEA), 1-Hydroxybenzotriazole (HOBT) and EDCI.
Further, the fluorine substituted Boc-L-phenylalanine in the step a1 has the structural formula
Figure GDA0004212129440000042
Represents that the F substitution position is any one of ortho position, meta position or para position, and the diglycolamine is +.>
Figure GDA0004212129440000043
Further, the molar ratio of the fluorine substituted Boc-L-phenylalanine to the diglycolamine, the N, N-diisopropylethylamine, the 1-hydroxybenzotriazole and the EDCI in the step a1 is 1:1.5:3.6:1.3:2.
Further, the volume ratio of dichloromethane to trifluoroacetic acid in the mixed solution of dichloromethane and trifluoroacetic acid in the step a2 is (1-4): 1.
Further, the structural formula of the terephthaloyl chloride in the step a3 is
Figure GDA0004212129440000044
The mol ratio of terephthaloyl chloride to L-NF-phe trifluoroacetate to triethylamine is 1 (2.1-2.3) to 10-15.
Further, the dosage of the sequential washing of the dichloromethane, the ethanol and the deionized water in the step a3 is respectively 30-50 mL, 10-20 mL and 30-50 mL.
(2) When the chiral hydrogel material with the functional halogenation effect is of a structural formula shown as II, the preparation steps are as follows:
b1, slowly dropwise adding thionyl chloride into a dry methanol solution under the protection of nitrogen and ice bath condition, slowly heating the reaction solution to room temperature after 10min, adding halogen atom-L-phenylalanine, reacting overnight at room temperature, and spin-drying to obtain methyl ester-protected halogenated L-phenylalanine, wherein the structural formula is as follows
Figure GDA0004212129440000051
X is any one of fluorine, chlorine, bromine or iodine;
b2, dissolving the methyl ester protected halogenated L-phenylalanine obtained in the step b1 in dichloromethane, cooling in an ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 24 hours, filtering the obtained colloid after the reaction is finished, sequentially washing dichloromethane, ethanol and deionized water, and drying the sample to obtain white solid L-4X-phe-Ome, wherein the structural formula is as follows
Figure GDA0004212129440000052
X is any one of fluorine, chlorine, bromine or iodine;
b3, dissolving the L-4X-phe-OMe obtained in the step b2 in methanol, adding 2M sodium hydroxide solution, reacting for 24 hours at room temperature, adjusting the PH to 2-3 by using 1M hydrochloric acid solution, filtering the colloid, washing with deionized water, and drying the sample to obtain white solid L-4X-phe-OH, wherein the structural formula is as follows
Figure GDA0004212129440000053
X is any one of fluorine, chlorine, bromine or iodine;
b4-1, dissolving the L-4X-phe-OH and the 1-hydroxybenzotriazole obtained in the step b3 with methylene dichloride, cooling in an ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, filtering the colloid, and filtering with dichloromethane, ethanol and deionized waterWashing and drying the sample sequentially to obtain a white solid target product II L-4X-phe, wherein the structural formula is
Figure GDA0004212129440000061
X is any one of fluorine, chlorine, bromine or iodine.
b4-2, dropwise adding 0.5mL of concentrated hydrochloric acid into the mixture of the L-4X-phe-OH obtained in the step b3 and 5 times of equivalent diethylene glycol, heating to 135 ℃, reacting for 4 hours, slowly dropwise adding deionized water after the reaction is finished, filtering the obtained colloid, washing the deionized water, and drying the sample to obtain a white solid target product IIL-4X-ES, wherein the structural formula is as follows
Figure GDA0004212129440000062
X is any one of fluorine, chlorine, bromine or iodine.
Further, when the chiral hydrogel material functionalized by the halogenation effect has the structural formula shown in II, the structural formula of the halogen atom-L-phenylalanine in the step b1 is
Figure GDA0004212129440000063
X is any one of fluorine, chlorine, bromine or iodine.
Further, the molar ratio of the halogen atom-L-phenylalanine to the thionyl chloride in the step b1 is 1:3.
Further, the structural formula of the terephthaloyl chloride in the step b2 is
Figure GDA0004212129440000064
The molar ratio of terephthaloyl chloride to methyl ester protected halogenated L-phenylalaninol and triethylamine is 1 (2.1-2.3) (10-15).
Further, the volume ratio of methanol to 2M sodium hydroxide solution in the step b3 is 3:1.
Further, the diglycolamine in the step b4-1 is structurally as follows
Figure GDA0004212129440000071
The molar ratio of L-4X-phe-OH to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:the dosage of the sequential washing of the methylene dichloride, the ethanol and the deionized water is 30-50 mL, 10-20 mL and 30-50 mL respectively according to the ratio of 3:7.2:2.6:4.
(3) When the chiral hydrogel material with the functional halogenation effect is shown in the structural formula III, the preparation steps are as follows:
and c1, slowly dropwise adding thionyl chloride into the dried methanol solution under the protection of nitrogen and ice bath condition, slowly heating the reaction solution to room temperature after 10min, adding halogen atom-L-phenylalanine, and reacting at room temperature for 12h. Spin-drying to obtain methyl ester protected halogenated L-phenylalanine
Figure GDA0004212129440000072
X is any one of fluorine, chlorine, bromine or iodine;
c2, dissolving the terephthalic acid and the 1-hydroxybenzotriazole with methylene dichloride, cooling in an ice bath, adding N, N-diisopropylethylamine and the halogenated L-phenylalanine protected by methyl ester obtained in the step c1, and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, suction-filtering the obtained colloid Buchner funnel, sequentially washing with dichloromethane, ethanol and deionized water, and oven drying the sample to obtain white solid L-4X-PA-OMe with the structural formula of
Figure GDA0004212129440000073
X is any one of fluorine, chlorine, bromine or iodine;
c3, dissolving the L-4X-PA-OMe obtained in the step c2 in methanol, adding 2M sodium hydroxide solution, reacting for 24 hours at room temperature, adjusting the pH to 2-3 by using 1M hydrochloric acid solution, filtering the obtained colloid Buchner funnel, washing with deionized water, and drying the sample in an oven to obtain white solid L-4X-PA-OH, wherein the structural formula is as follows
Figure GDA0004212129440000081
X is any one of fluorine, chlorine, bromine or iodine;
c4, dissolving the L-4X-PA-OH obtained in the step c3 and 1-hydroxybenzotriazole with methylene dichloride, cooling in an ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, suction-filtering the obtained colloid Buchner funnel, sequentially washing with dichloromethane, ethanol and deionized water, and drying the sample in an oven to obtain a white solid target product III L-4X-PA, wherein the structural formula is
Figure GDA0004212129440000082
X is any one of fluorine, chlorine, bromine or iodine.
Further, when the chiral hydrogel material functionalized by the halogenation effect has the structural formula shown in III, the structural formula of the halogen atom-L-phenylalanine in the step c1 is
Figure GDA0004212129440000083
X is any one of fluorine, chlorine, bromine or iodine.
Further, the molar ratio of the halogen atom-L-phenylalanine to the thionyl chloride in the step c1 is 1:3.
Further, the halogen atom substituted L-phenylalanine in the step c1 is subjected to esterification reaction in the presence of thionyl chloride and methanol to produce methyl ester protected halogenated L-phenylalanine.
Further, the molar ratio of the halogenated L-phenylalanine protected by methyl ester to the terephthaloyl acetic acid, the N, N-diisopropylethylamine, the 1-hydroxybenzotriazole and the EDCI in the step c2 is 2:1:7.2:2.6:4, and the consumption of the sequential washing of the dichloromethane, the ethanol and the deionized water is 30-50 mL, 10-20 mL and 30-50 mL respectively.
Further, in step c2, methyl ester protected halogenated L-phenylalanine and terephthaloyl acid undergo an amide condensation reaction in the presence of N, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI to produce L-4X-PA-OMe.
Further, the volume ratio of methanol to 2M sodium hydroxide solution in the step c3 is 3:1.
Further, in step c3, L-4X-PA-OMe is de-methyl in the presence of methanol, 2M sodium hydroxide solution, yielding L-4X-PA-OH.
Further, the structure of the diglycolamine in the step c4 is
Figure GDA0004212129440000091
The molar ratio of L-4X-PA-OH to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:3:7.2:2.6:4.
Furthermore, the chiral hydrogel material functionalized by the halogenation effect is applied to promotion of cell adhesion and proliferation through molecular chirality and assembly fiber chirality.
Further, the cells are Hep G2 cells.
Further, the cells are NIH 3T3 cells.
The invention has the following beneficial effects:
1. compared with the traditional method of reversing the chiral of the spiral structure caused by enantiomer molecules, solvents, temperature, guest molecules, illumination, pH and other variables, the invention obviously improves the defects of poor biocompatibility, unstable structure, difficult operation and the like of the traditional system.
2. The chiral nanofiber material constructed by single enantiomer regulates cell adhesion behavior through the change of molecular chirality and assembly fiber chirality, and the effects of the molecular chirality and supermolecular chirality in the process of regulating cell behavior by chiral nanofibers are disclosed.
3. The halogenation effect functionalized chiral hydrogel material prepared by the invention has the advantages of simple synthesis, good biocompatibility and degradability, is a chiral nanofiber constructed by a single enantiomer, is easy to regulate and control in chirality, and is expected to have unique application in the fields of cell tissue culture and other medicine.
4. The preparation method of the chiral hydrogel material with the functionalized halogenation effect does not need complex reaction, is easy for large-scale production, and has high purity of the obtained product; the adhesion behavior of the chiral nanofiber hydrogel regulatory cells is carried out in a physiological environment, so that the chiral nanofiber hydrogel regulatory cell is convenient to operate and has practical application value.
5. When the chiral hydrogel material with the functional halogenation effect prepared by the invention is applied to cell adhesion and proliferation, especially adhesion and proliferation of Hep G2 cells and NIH 3T3 cells, the number of NIH 3T3 cells adhered to an L-4F-phe (LM) nanofiber membrane is 2.2 times that of the number of NIH 3T3 cells adhered to a D-4F-phe (DP) membrane, 1.6 times that of the NIH 3T3 cells adhered to the L-phe (LP) membrane, 1.3 times that of the NIH 3T3 cells adhered to a D-phe (DM) membrane, and 1.75 times that of the NIH 3T3 cells adhered to a PS pore. The number of cells adhered to the L-4F-phe (LM) membrane by HepG2 cells was 1.9 times, 1.3 times, 1.1 times and 1.5 times the number of cells on the D-4F-phe (DP) membrane, on the L-phe (LP) membrane, on the D-phe (DM) membrane and on the PS well, respectively.
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In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a photograph of a chiral hydrogel material functionalized by halogenation effect prepared in example 1 of the present invention, wherein A is a photograph of a hydrogel functionalized by substitution of fluorine atoms at different positions, and B and C are photographs of a hydrogel functionalized by different halogen atoms;
FIG. 2 is a graph showing comparison of nanofiber scans of L-phe, L-2F-phe, L-3F-phe, and L-4F-phe of the supramolecular hydrogels prepared in example 1 of the present invention; wherein, L-phe and L-2F-phe are right-handed helices, L-3F-phe and L-4F-phe are left-handed helices.
FIG. 3 is a fluorescence micrograph and cell adhesion density of NIH 3T3 cells and HepG2 cells of the invention after incubation on halogenated effect functionalized chiral nanofibers L-phe, L-4F-phe, D-phe and D-4F-phe films of the invention for various times; wherein, A) is the quantitative data of the L-Phe, L-4F-Phe, D-Phe and D-4F-Phe nanofiber membranes and NIH 3T3 cells of the PS control plate after 3 days of culture and 1 day of incubation, 2 days, and 3 days later, the L-Phe, L-4F-Phe, D-Phe and D-4F-Phe nanofiber membranes and the NIH 3T3 cells of the PS control, N=6; * Data shows significant differences (ANOVA: p0.05,: p0.005,: p 0.001); b) Quantitative data for HepG2 cells of L-Phe, L-4F-Phe, D-Phe and D-4F-Phe nanofiber membranes and PS controls after 3 days of incubation and 1 day, 2 days, 3 days of incubation, n=6; * Data shows significant differences (ANOVA: p0.05,: p0.005,: p 0.001); c) Confocal microscopy pictures of three-dimensional cells of chiral hydrogels after 3 days of culture.
FIG. 4 is a circular dichroism spectrum of the fluorinated functionalized chiral hydrogel material prepared in example 1 of the present invention.
FIG. 5 shows a chiral hydrogel material L-phe prepared in example 1 of the present invention 1 HNMR spectra.
FIG. 6 shows a fluorinated effect functionalized chiral hydrogel material L-2F-phe prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 7 shows a fluorinated effect functionalized chiral hydrogel material L-3F-phe prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 8 shows a fluorinated effect functionalized chiral hydrogel material L-4F-phe prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 9 is a schematic illustration of a chiral hydrogel material L-4Cl-phe functionalized by the halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 10 shows a chiral hydrogel material L-4Br-phe functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 11 shows a chiral hydrogel material L-4I-phe functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 12 is a schematic illustration of a chiral hydrogel material L-4F-ES functionalized with halogenated effects prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 13 is a schematic illustration of a chiral hydrogel material L-4Cl-ES functionalized by the halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 14 shows a chiral hydrogel material L-4Br-ES functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 15 is a schematic illustration of the functionalization of the halo-effect prepared in example 1 of the present inventionChiral hydrogel material L-4I-ES 1 H NMR spectrum.
FIG. 16 shows a chiral hydrogel material L-4F-PA functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 17 is a schematic illustration of a halo-effect functionalized chiral hydrogel material L-4Cl-PA prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 18 shows a chiral hydrogel material L-4Br-PA functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
FIG. 19 is a schematic diagram showing a chiral hydrogel material L-4I-PA functionalized by halogenated effect prepared in example 1 of the present invention 1 H NMR spectrum.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without any inventive effort, are intended to be within the scope of the invention.
Example 1
The embodiment is a chiral uniform supermolecular gel nanofiber material for cell adhesion culture, the chiral material changes the spatial conformation of a gel factor by utilizing different halogen atoms and substitution positions thereof or the hydrophilic and hydrophobic changes of side chains, the gel factor constructed by single enantiomer is self-assembled into different chiral nanofiber materials through physical interaction, the gel factors are L-or D-phenylalanine derivatives, and the molecular structural formula is as follows:
Figure GDA0004212129440000121
wherein R1 in part a is H or NH 2 (CH 2 ) 2 O(CH 2 ) 2 In the OH, L-phenylalanine derivatives, the substitution positions of F are respectively ortho (L-2F-phe), meta (L-3F-phe) and para (L-4F-phe); the D-phenylalanine derivative is para (D-4F-phe).
In part b, L-phenylalanine derivatives, wherein R is NH 2 (CH 2 ) 2 O(CH 2 ) 2 OH (L-4X-phe) or HO (CH) 2 ) 2 O(CH 2 ) 2 OH (L-4X-ES); x is F, cl, br, I.
In part c, L-phenylalanine derivatives, wherein R1 is NH 2 (CH 2 ) 2 O(CH 2 ) 2 OH (L-4X-PA); x is F, cl, br, I.
In the part a, chiral inversion controllability of the supermolecule assembly is realized by utilizing different substitution positions of fluorine atoms. In the part b, the chiral inversion of the supermolecule assembly is controllable by utilizing the substitution difference of different halogen atoms. In the part c, the chiral inversion of the supermolecule assembly is controllable by utilizing the substitution difference of different halogen atoms. In the self-assembly process of the chiral gel factor, a molecular chiral signal is transmitted to a nanofiber assembly structure to obtain the chiral gel fiber similar to the collagen spiral structure in a human body. The L-phenylalanine derivative of the a part can obtain right-handed spiral nano fiber when fluorine atoms are in ortho positions (L-2F-phe); when the fluorine atom is in meta position (L-3F-phe) and para position (L-4F-phe), the left-handed spiral nanofiber can be obtained; the D-phenylalanine derivative part can obtain right-handed spiral nano fiber when fluorine atoms are in para position (D-4F-phe).
The preparation method of the chiral hydrogel material with the halogenated effect comprises the following steps:
(1) a step of synthesizing a part of gel factor:
step a1, preparation of Boc-L-NF-phe
Figure GDA0004212129440000131
Preparation of Boc-L-NF-phe: dissolving fluorine-substituted Boc-L-phenylalanine and 1-hydroxybenzotriazole with dichloromethane, cooling in ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, and continuing the room temperature reaction for 12 hours; sequentially washing with saturated citric acid solution, saturated sodium bicarbonate solution and saturated sodium chloride solution, spin drying, and recrystallizing with ethyl acetate and petroleum ether, wherein the molar ratio of fluorine-substituted Boc-L-phenylalanine to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:1.5:3.6:1.3:2.
Step a2, synthesis of L-NF-phe trifluoroacetate salt
Figure GDA0004212129440000132
Boc-L-NF-phe is added into a solution with the volume ratio of dichloromethane/trifluoroacetic acid of 4/1, and after reaction for 3 hours at room temperature, the solution is subjected to rotary evaporation to completely remove the solution and the residual trifluoroacetic acid of L-NF-phe.
Step a3, preparing gel factors at different substitution positions of fluorine atoms:
Figure GDA0004212129440000133
dissolving the trifluoroacetate of the L-NF-phe in dichloromethane, cooling in an ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, and reacting at room temperature for 24 hours. Filtering the obtained colloid Buchner funnel, sequentially washing dichloromethane, ethanol and deionized water, and drying a sample in an oven to obtain a white solid target product I, wherein the molar ratio of terephthaloyl chloride to L-NF-phe trifluoroacetate to triethylamine is 1:2.2:10
According to the substitution positions of fluorine, o-, m-and p-fluorine substituted Boc-L-phenylalanine can be selected, so that L-2F-phe, L-3F-phe and L-4F-phe can be prepared respectively according to the above conditions.
The fluorine para-substituted Boc-D-phenylalanine was selected, whereby D-4F-phe was produced according to the above conditions.
(2) And b, partial gel factor synthesis:
step b1, synthesis of methyl ester protected halogenated L-phenylalanine
Figure GDA0004212129440000141
Under the protection of nitrogen and ice bath, thionyl chloride is slowly dripped into the dried methanol solution, after 10min, the reaction solution is slowly warmed to room temperature, halogen atom-L-phenylalanine is added, and the reaction is carried out for 12h at room temperature. The next reaction can be carried out after spin drying, and the molar ratio of halogen atom-L-phenylalanine to thionyl chloride is 1:3.
Step b2, synthesis of L-4X-phe-OMe
Figure GDA0004212129440000142
Dissolving methyl ester protected halogenated L-phenylalanine in dichloromethane, cooling in ice bath, sequentially adding triethylamine, terephthaloyl chloride and terephthaloyl chloride, slowly heating the reaction solution to room temperature in a molar ratio of 1:2.2:10, stirring, and reacting at room temperature for 24 hours. The obtained colloid buchner funnel is filtered, 100mL of dichloromethane, 100mL of ethanol and 500mL of deionized water are washed in sequence, and a sample is dried in an oven to obtain white solid L-4X-phe-OMe.
Step b3, preparation of L-4X-phe-OH:
Figure GDA0004212129440000143
dissolving L-4X-phe-OMe in methanol, adding 2M sodium hydroxide solution, reacting the methanol with the 2M sodium hydroxide solution at a volume ratio of 3:1 at room temperature for 24 hours, adjusting the PH to 2-3 by using 1M hydrochloric acid solution, carrying out suction filtration on the obtained colloid buchner funnel, washing with deionized water, and drying the sample in an oven to obtain white solid L-4X-phe-OH.
Step b4, preparation of different halogen atom functionalized gelators:
Figure GDA0004212129440000151
b 4-1) dissolving L-4X-phe-OH and 1-hydroxybenzotriazole with methylene dichloride, cooling in an ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; slowly raising the reaction solution to room temperature, stirring, reacting at room temperature for 12 hours, carrying out suction filtration on the obtained colloid Buchner funnel, sequentially washing dichloromethane, ethanol and deionized water, and drying a sample in an oven to obtain a white solid target product II L-4X-phe, wherein the molar ratio of L-4X-phe-OH to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:3:7.2:2.6:4.
When (when)
Figure GDA0004212129440000152
When the halogen atoms are Cl, br and I respectively, the Cl-substituted, br-substituted and I-substituted L-phenylalanine can be selected according to the difference of the halogen atoms on the para position of the halogen atom-L-phenylalanine, so that the L-4Cl-phe, the L-4Br-phe and the L-4I-phe can be prepared respectively according to the conditions.
b 4-2) dropwise adding concentrated hydrochloric acid into the mixture of L-4X-phe-OH and diethylene glycol, heating to 135 ℃, reacting for 4 hours, slowly dropwise adding deionized water after the reaction is finished, filtering the obtained colloid, washing the deionized water, and drying a sample in an oven to obtain a white solid target product IIL-4X-ES.
When (when)
Figure GDA0004212129440000153
When the halogen atoms are F, cl, br, I, L-phenylalanine substituted with F, cl, br and I can be selected according to the difference of the halogen atoms at the para position of the halogen atom-L-phenylalanine, thereby preparing L-4F-ES, L-4Cl-ES, L-4Br-ES and L-4I-ES respectively according to the above conditions.
(3) c, partial gel factor synthesis:
step c1, synthesis of methyl ester protected halogenated L-phenylalanine
Figure GDA0004212129440000154
Under the protection of nitrogen and ice bath, thionyl chloride is slowly added into a dry methanol solution in a dropwise manner, after 10min, the reaction solution is slowly warmed to room temperature, halogen atom-L-phenylalanine is added, and the molar ratio of the halogen atom-L-phenylalanine to the thionyl chloride is 1:3 for reaction for 12 hours at room temperature. Spin drying to perform the next reaction.
Step c2, synthesis of L-4X-PA-OMe
Figure GDA0004212129440000161
Dissolving terephthalic acid and 1-hydroxybenzotriazole with dichloromethane, cooling in ice bath, adding N-diisopropylethylamine and methyl ester-protected halogenated L-phenylalanine, ice-bathing for 30min, and adding EDCI; the reaction solution is slowly warmed to room temperature and stirred, the reaction is carried out for 12 hours at room temperature, the obtained colloid buchner funnel is filtered, dichloromethane, ethanol and deionized water are sequentially washed, a sample is dried in an oven, and the white solid L-4X-PA-Ome, methyl ester protected halogenated L-phenylalanine and terephthalic acid, N-diisopropylethylamine and 1-hydroxybenzotriazole are obtained, wherein the molar ratio of EDCI is 2:1:7.2:2.6:4.
Step c3, preparation of L-4X-PA-OH:
Figure GDA0004212129440000162
dissolving L-4X-PA-OMe in methanol, adding 2M sodium hydroxide solution, reacting methanol and 2M sodium hydroxide solution at a volume ratio of 3:1 at room temperature for 24 hours, adjusting pH to 2-3 with 1M hydrochloric acid solution, filtering the obtained colloid Buchner funnel, washing with deionized water, and drying the sample in an oven to obtain white solid L-4X-PA-OH.
Step c4, preparing different halogen atom functionalized gel factors:
Figure GDA0004212129440000163
dissolving the L-4X-PA-OH and the 1-hydroxybenzotriazole with methylene dichloride, cooling in an ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; the molar ratio of L-4X-PA-OH to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:3:7.2:2.6:4, the reaction solution is slowly warmed to room temperature and stirred, the reaction is carried out at room temperature for 12 hours, the obtained colloid Buchner funnel is filtered, dichloromethane, ethanol and deionized water are sequentially washed, and a sample is dried in an oven to obtain a white solid target product L-4X-PA.
According to the difference of halogen atoms on the para position of the halogen atom-L-phenylalanine, when the halogen atoms are F, cl, br, I, the L-phenylalanine with F substitution, cl substitution, br substitution and I substitution can be selected, so that L-4F-PA, L-4Cl-PA, L-4Br-PA and L-4I-PA can be respectively prepared according to the above conditions.
Comparative example 1
The preparation method of the comparative example L-phe comprises the following steps:
step a1, preparation of Boc-L-N-phe
Figure GDA0004212129440000171
Preparation of Boc-L-N-phe: dissolving Boc-L-phenylalanine and 1-hydroxybenzotriazole in dichloromethane, cooling in ice bath, adding diglycolamine and N, N-diisopropylethylamine, ice bath for 30min, and adding EDCI; slowly heating the reaction solution to room temperature, and continuing the room temperature reaction for 12 hours; sequentially washing with saturated citric acid solution, saturated sodium bicarbonate solution and saturated sodium chloride solution, spin drying, and recrystallizing with ethyl acetate and petroleum ether, wherein the molar ratio of Boc-L-phenylalanine to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:1.5:3.6:1.3:2.
Step a2, synthesis of L-N-phe trifluoroacetate salt
Figure GDA0004212129440000172
Boc-L-NF-phe was added to a solution of 4/1 methylene chloride/trifluoroacetic acid (V/V), and after 3 hours of reaction at room temperature, the solution was rotary evaporated to completely remove the solution and the remaining L-N-phe of trifluoroacetic acid.
Step a3, preparation of gelator
Figure GDA0004212129440000173
Dissolving the trifluoroacetate of the L-N-phe in dichloromethane, cooling in an ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, and reacting at room temperature for 24 hours. And (3) carrying out suction filtration on the obtained colloid Buchner funnel, sequentially washing with dichloromethane, ethanol and deionized water, and drying a sample in an oven to obtain a target product L-phe, wherein the molar ratio of terephthaloyl chloride to L-N-phe trifluoroacetate to triethylamine is 1:2.2:10.
Comparative example 2
The preparation method of the comparative example D-phe comprises the following steps:
step a1, preparation of Boc-D-N-phe
Preparation of Boc-D-N-phe: dissolving Boc-D-phenylalanine and 1-hydroxybenzotriazole in dichloromethane, cooling in ice bath, adding diglycolamine and N, N-diisopropylethylamine, ice bath for 30min, and adding EDCI; slowly heating the reaction solution to room temperature, and continuing the room temperature reaction for 12 hours; sequentially washing with saturated citric acid solution, saturated sodium bicarbonate solution and saturated sodium chloride solution, spin drying, and recrystallizing with ethyl acetate and petroleum ether, wherein the molar ratio of Boc-D-phenylalanine to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:1.5:3.6:1.3:2.
Step a2, synthesis of D-N-phe trifluoroacetate salt
Boc-D-N-phe was added to a solution of 4/1 methylene chloride/trifluoroacetic acid (V/V) and after 3h reaction at room temperature, the solution was rotary evaporated to remove the solution and the remaining D-N-phe of trifluoroacetic acid completely.
Step a3, preparation of gelator
Dissolving the trifluoroacetate of the D-N-phe in dichloromethane, cooling in an ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, and reacting at room temperature for 24 hours. And (3) carrying out suction filtration on the obtained colloid Buchner funnel, sequentially washing with dichloromethane, ethanol and deionized water, and drying a sample in an oven to obtain a target product D-phe.
The halogenated effect functionalized supramolecular hydrogels prepared in this example are shown in fig. 1A, 1B and 1C: FIG. 1A is a fluorinated functionalized hydrogel, L-2F-phe, L-3F-phe and L-4F-phe, respectively; FIG. 1B is a schematic diagram of a hydrogel functionalized with different halogen atoms, L-4Cl-phe, L-4Br-phe, L-4I-phe, L-4F-ES, L-4Cl-ES, L-4Br-ES, and L-4I-ES, respectively; FIG. 1C shows hydrogels functionalized with different halogen atoms, L-4F-PA, L-4Cl-PA, L-4Br-PA, and L-4I-PA, respectively. Gel factor of chiral Structure functionalized by halogenation Effect prepared in example 1 1 The H nuclear magnetic spectrum is shown in figures 5-12 in sequence, and the L-phe is shown in figure 5 1 H NMR spectrum, FIG. 6 is L-2F-phe 1 HNMR spectra, FIG. 7 is L-3F-phe 1 H NMR spectrum, FIG. 8 is L-4F-phe 1 H NMR spectrum, FIG. 9 is L-4Cl-phe 1 H NMR spectrum, FIG. 10 is L-4Br-phe 1 HNMR spectra, FIG. 11 is L-4I-phe 1 HNMR spectra, FIG. 12 is L-4F-ES 1 HNMR spectra, FIG. 13 is L-4Cl-ES 1 H NMR spectrum, FIG. 14 is L-4Br-ES 1 H NMR spectrum, FIG. 15 is L-4I-ES 1 H NMR spectrum, FIG. 16 is L-4F-PA 1 H NMR spectrum, FIG. 17 is L-4Cl-PA 1 H NMR spectrum, FIG. 18 is L-4Br-PA 1 H NMR spectrum, FIG. 19 is L-4I-PA 1 HNMR spectra.
Characterization analysis was performed on L-phe and fluorinated effect functionalized gels L-2F-phe, L-3F-phe and L-4F-phe microtopography using Scanning Electron Microscopy (SEM). As shown in fig. 2, a) L-phe, b) L-2F-phe, c) L-3F-phe, d) L-4F-phe, respectively, wherein L-phe and L-2F-phe form right-handed helical fibers and L-3F-phe and L-4F-phe form left-handed helical fibers.
Circular Dichroism (CD) was further used to verify that the fluorination effect induced inversion of supramolecular chirality, as shown in figure 4. In FIG. 4a, the CD spectrum of hydrogel L-phe shows a pronounced Ketone effect, zero crossing at 264nm, a slightly weaker positive peak at 303nm, and a stronger negative peak at 208 nm; similarly, in FIG. 4b, a positive effect peak, which is stronger at 278nm and a negative effect peak, which is weaker at 223nm, of the hydrogel L-2F-phe are observed; as expected, the Ketone effect of L-3F-phe and L-4F-phe was reversed in the high wave region compared to the gels L-phe and L-2F-phe, L-3F-phe had two negative effect peaks at 229nm and 315nm, and L-4F-phe had two negative effect peaks at 230nm and 300 nm. The results are very consistent with SEM results.
Application example: use of fluorinated effect functionalized chiral nanofiber material in cell adhesion and proliferation
HepG2 cells and NIH 3T3 cells used in the present invention were purchased from Shanghai Shaoxing Biotechnology Co.
Adhesion amount regulation of HepG2 cells and NIH 3T3 cells on surface of chiral uniform nanofiber membrane functionalized by fluorination effect
The nanofiber membrane and the three-dimensional hydrogel adopted in the application example are prepared by self-assembling phenylalanine derivative gel factors with uniform chirality, which are prepared in the previous example 1.
For the two-dimensional fiber film, various gel factors are firstly prepared into 0.3mg/mL of ultrapure water dilute solution respectively, cooled and self-assembled, 100 mu L of 0.3mg/mL of hydrogel dilute solution is added into each hole of a 96-hole cell culture plate, vacuum drying is carried out for 12 hours at 37 ℃, ultraviolet light irradiation is carried out for 30 minutes for sterilization, NIH 3T3 cells and HepG2 cells are inoculated on the 96-hole cell culture plate (about 2000 holes) coated with the functionalized xerogel film, and the 96-hole cell culture plate is placed at 37 ℃ and 5 percent CO 2 Incubation in a cell incubator was performed to observe cell adhesion at various time points. Live-dead cell double staining is adopted, observation and photographing are carried out under a fluorescence microscope, and statistical analysis is carried out by using Image J software, and the chart is shown in figure 3; as can be seen from FIGS. 3A and 3B, the NIH 3T3 cells adhered to the L-4F-phe (LM) nanofiber membrane in an amount of 2.2 times that on the D-4F-phe (DP) membrane, 1.6 times that on the L-phe (LP) membrane, 1.3 times that on the D-phe (DM) membrane, and 1.75 times that on the PS wells. The number of cells adhering to the L-4F-phe (LM) membrane by HepG2 cells was 1.9 times, 1.3 times, 1.1 times and 1.5 times the number of cells on the D-4F-phe (DP) membrane, on the L-phe (LP) membrane, on the D-phe (DM) membrane and on the PS well, respectively.
For the three-dimensional hydrogel, firstly, various gel factors are respectively prepared into DMSO mother solution, and then NIH 3T3 cell suspension cells are mixed into DMSO #Final DMSO concentration: 5%) in the gel concentrate (final gel concentration: 10 mg/mL), a cell-encapsulated hydrogel was formed in a few minutes, more DMEM was then added to the hydrogel and transferred to standard culture conditions (37 ℃,5% co) 2 ) Is contained in a humidified incubator. After 3 days of culture, the cells were uniformly distributed in the hydrogel and did not sediment (fig. 3C). As can be seen from the green fluorescence emitted from living cells, the L-4F-phe (LM) hydrogel had a higher cell density than the D-4F-phe (DP), L-phe (LP) and D-phe (DM) hydrogels.
In conclusion, the nano gel material with uniform chirality prepared by the implementation successfully imitates the chiral spiral structure of the extracellular microenvironment, and the molecular chirality and the assembled fiber chirality can obviously influence the adhesion growth behavior of cells. Studies have shown that the effect of Gao Jiechao molecular chirality on cell proliferation and viability is more pronounced than that of monomer chirality, and that the positive interaction order of supermolecular chirality and monomer chirality on cell proliferation and viability is LM > DM > LP > DP.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.

Claims (10)

1. A chiral hydrogel material functionalized by halogenation effect, characterized in that: the structural formula of the chiral hydrogel material with the functional halogenation effect is as follows
Figure QLYQS_1
F represents any one of ortho, meta or para positions of F substitution.
2. A chiral hydrogel material functionalized by halogenation effect, characterized in that: the structural formula of the chiral hydrogel material with the functional halogenation effect is as follows
Figure QLYQS_2
Or (b)
Figure QLYQS_3
X is any one of fluorine, chlorine, bromine or iodine.
3. A chiral hydrogel material functionalized by halogenation effect, characterized in that: the structural formula of the chiral hydrogel material with the functional halogenation effect is as follows
Figure QLYQS_4
X is any one of fluorine, chlorine, bromine or iodine.
4. The method for preparing the chiral hydrogel material with the functionalized halogenation effect according to claim 1, which is characterized by comprising the following preparation steps:
a1 Boc-LDissolving phenylalanine and 1-hydroxybenzotriazole in dichloromethane, cooling in ice bath, adding diglycolamine and N, N-diisopropylethylamine, and adding EDCI after ice bath for 30 min; slowly heating the reaction solution to room temperature, and continuing the reaction at room temperature overnight; sequentially washing with saturated citric acid solution, saturated sodium bicarbonate solution, saturated sodium chloride solution, spin drying, and recrystallizing with ethyl acetate and petroleum ether to obtain Boc-LNF-phe of the formula
Figure QLYQS_5
-F represents any one of ortho, meta or para positions with respect to the F substitution position; the fluorine-substituted Boc-in step a1L-phenylalanine of the formula +>
Figure QLYQS_6
-F represents any one of ortho, meta or para positions with respect to the F substitution position;
a2, boc-LAdding NF-phe into a mixed solution of dichloromethane and trifluoroacetic acid, reacting for 3 hours at room temperature, and removing Boc-protecting group to obtainLNF-phe trifluoroacetate with the structural formula
Figure QLYQS_7
-F represents any one of ortho, meta or para positions with respect to the F substitution position;
a3, the step a2 is carried outLDissolving NF-phe trifluoroacetate in dichloromethane, cooling in ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 24h, filtering the colloid after the reaction, sequentially washing dichloromethane, ethanol and deionized water, and drying the sample to obtain a target product I, wherein the structural formula is
Figure QLYQS_8
F represents any one of ortho, meta or para positions of F substitution.
5. The method for preparing the chiral hydrogel material functionalized by halogenation effect according to claim 4, wherein: the fluorine substituted Boc-L-phenylalanine to diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole, EDCI in a molar ratio of 1:1.5:3.6:1.3:2; the volume ratio of the dichloromethane to the trifluoroacetic acid in the mixed solution of the dichloromethane and the trifluoroacetic acid in the step a2 is (1-4): 1; the terephthaloyl chloride in the step a3LThe mol ratio of the NF-phe trifluoroacetate to the triethylamine is 1 (2.1-2.3) (10-15).
6. The method for preparing the chiral hydrogel material with the functionalized halogenation effect according to claim 2, which is characterized by comprising the following preparation steps:
b1, slowly dropwise adding thionyl chloride into a dry methanol solution under the protection of nitrogen and ice bath condition, slowly heating the reaction solution to room temperature after 10min, and adding halogen atoms-LPhenylalanine, overnight reaction at room temperature, spin-drying to obtain methyl ester protected halogenatedLPhenylalanine of the formula
Figure QLYQS_9
X is any one of fluorine, chlorine, bromine or iodine; halogen atom in the step b 1-L-phenylalanine of the formula +.>
Figure QLYQS_10
X is any one of fluorine, chlorine, bromine or iodine;
b2, methyl ester-protected halogenated obtained in step b1LDissolving phenylalanine in dichloromethane, cooling in ice bath, sequentially adding triethylamine and terephthaloyl chloride, slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 24h, filtering the colloid after the reaction, sequentially washing with dichloromethane, ethanol and deionized water, and drying to obtain solidL-4X-phe-OMe of the formula
Figure QLYQS_11
X is any one of fluorine, chlorine, bromine or iodine;
b3, the step b2 is carried outLdissolving-4X-phe-OMe in methanol, adding 2M sodium hydroxide solution, reacting at room temperature for 24h, adjusting pH to 2-3 with 1M hydrochloric acid solution, suction filtering the colloid, washing with deionized water, and drying to obtain solidL-4X-phe-OH of the formula
Figure QLYQS_12
X is any one of fluorine, chlorine, bromine or iodine;
b4-1, the step b3L-4X-phe-OH and 1-hydroxybenzotriazole are dissolved in methylene dichloride, cooled in an ice bath, diglycolamine and N, N-diisopropylethylamine are added, and EDCI is added after ice bath for 30 min; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, filtering the colloid after the reaction is finished, washing dichloromethane, ethanol and deionized water in sequence, and drying the sample to obtain a target productL-4X-phe of the formula
Figure QLYQS_13
X is any one of fluorine, chlorine, bromine or iodine;
b4-2, to step b3LDropwise adding 0.5mL of concentrated hydrochloric acid into a mixture of 4X-phe-OH and 5 times of diethylene glycol, heating to 135 ℃, reacting for 4 hours, slowly dropwise adding deionized water after the reaction is completedFiltering the colloid, washing with deionized water, and drying the sample to obtain the target productL-4X-ES of the formula
Figure QLYQS_14
X is any one of fluorine, chlorine, bromine or iodine.
7. The method for preparing the chiral hydrogel material functionalized by halogenation effect according to claim 6, wherein: the halogen atom-L-phenylalanine to thionyl chloride molar ratio of 1:3; the terephthaloyl chloride and methyl ester protected halo in step b2LThe molar ratio of the phenylalaninol to the triethylamine is 1 (2.1-2.3) (10-15); the volume ratio of the methanol to the 2M sodium hydroxide solution in the step b3 is 3:1; in the step b4-1LThe molar ratio of the-4X-phe-OH to the diglycolamine, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI is 1:3:7.2:2.6:4.
8. A method for preparing a chiral hydrogel material functionalized for halogenation effect as claimed in claim 3, characterized by the following steps:
c1, slowly dropwise adding thionyl chloride into a dry methanol solution under the protection of nitrogen and under the ice bath condition, slowly heating the reaction solution to room temperature after 10min, and adding halogen atoms-LReacting phenylalanine at room temperature for 12h, and spin-drying to obtain methyl ester protected halogenated L-phenylalanine, wherein the structural formula is
Figure QLYQS_15
X is any one of fluorine, chlorine, bromine or iodine; halogen atom in the step c 1-L-phenylalanine of the formula +.>
Figure QLYQS_16
X is any one of fluorine, chlorine, bromine or iodine;
c2, dissolving the terephthaloyl acid and the 1-hydroxybenzotriazole with methylene dichloride, cooling in an ice bath, adding N, N-diisopropylethylamine and methyl ester protected halogenated obtained in the step c1L-phenylalanine, after 30min ice bath, EDCI was added; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, suction-filtering the obtained colloid, washing with dichloromethane, ethanol and deionized water in sequence, and drying the sample in an oven to obtain a white solidL-4X-PA-OMe of formula
Figure QLYQS_17
X is any one of fluorine, chlorine, bromine or iodine;
c3, the step c2 is carried outLdissolving-4X-PA-OMe in methanol, adding 2M sodium hydroxide solution, reacting at room temperature for 24h, adjusting pH to 2-3 with 1M hydrochloric acid solution, filtering, washing with deionized water, and oven drying to obtain solidL-4X-PA-OH of the formula
Figure QLYQS_18
X is any one of fluorine, chlorine, bromine or iodine;
c4, the step c3 is carried outL-4X-PA-OH and 1-hydroxybenzotriazole are dissolved in methylene dichloride, cooled in ice bath, diglycolamine and N, N-diisopropylethylamine are added, and EDCI is added after ice bath for 30 min; slowly heating the reaction solution to room temperature, stirring, reacting at room temperature for 12h, suction-filtering the obtained colloid, washing dichloromethane, ethanol and deionized water in sequence, and drying the sample in an oven to obtain a target productL-4X-PA of the formula
Figure QLYQS_19
X is any one of fluorine, chlorine, bromine or iodine.
9. The method for preparing the chiral hydrogel material functionalized by halogenation effect according to claim 8, wherein: halogen atom-L-phenylalanine to thionyl chloride molar ratio of 1:3; the molar ratio of the halogenated L-phenylalanine protected by methyl ester to the terephthaloyl acetic acid, N-diisopropylethylamine, 1-hydroxybenzotriazole and EDCI in the step c2 is 2:1:7.2:2.6:4; the volume ratio of the methanol to the 2M sodium hydroxide solution in the step c3 is 3:1; in the step c4L-4X-PA-OHThe mol ratio of diglycolamine to N, N-diisopropylethylamine to 1-hydroxybenzotriazole to EDCI is 1:3:7.2:2.6:4.
10. Use of the halogenation effect functionalized chiral hydrogel material according to claim 1 or 2 or 3 for the preparation of a medicament for promoting cell adhesion and proliferation by molecular chirality and assembly fiber chirality, characterized in that: the cells are NIH 3T3 cells or Hep G2 cells.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR3007760A1 (en) * 2013-06-28 2015-01-02 Arkema France FATTY AMIDE BASIC CAPROLACTAM AS GELLER ADDITIVE.
CN109569445A (en) * 2018-10-09 2019-04-05 上海交通大学 The chiral hydrogel material and preparation method of odd-even effect functionalization
CN112641995A (en) * 2020-12-21 2021-04-13 上海交通大学 Chiral hydrogel dressing with antibacterial and repair promoting functions and preparation method and application thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160002457A1 (en) * 2013-02-20 2016-01-07 Celluforce Inc. Tunable and responsive photonic hydrogels comprising nanocrystalline cellulose

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR3007760A1 (en) * 2013-06-28 2015-01-02 Arkema France FATTY AMIDE BASIC CAPROLACTAM AS GELLER ADDITIVE.
CN109569445A (en) * 2018-10-09 2019-04-05 上海交通大学 The chiral hydrogel material and preparation method of odd-even effect functionalization
CN112641995A (en) * 2020-12-21 2021-04-13 上海交通大学 Chiral hydrogel dressing with antibacterial and repair promoting functions and preparation method and application thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Helicity Inversion of Supramolecular Hydrogels Induced by Achiral Substituents;Guofeng Liu et al.;《ACS Nano》;第11卷;第11880−11889页 *
The Cooperative Effect of Both Molecular and Supramolecular Chirality on Cell Adhesion;Jinying Liu et al.;《Angew. Chem. Int. Ed》;第57卷;第6475-6479页 *
手性超分子凝胶材料的研究进展;刘国锋 等;《中国材料进展》;第32卷(第7期);第420-441页 *
水凝胶医用敷料的研究进展;樊梦妮 等;《生物加工过程》;第19卷(第3期);第294-295页 *

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